Regulation of glucose transport by interleukin-3 in growth factor-dependent and oncogene-transformed bone marrow-derived cell lines

doi:10.1016/S0145-2126(97)00016-7

Leukemia Research

Volume 21, Issue 7, July 1997, Pages 609-618

https://doi.org/10.1016/S0145-2126(97)00016-7 Get rights and content

Abstract

Growth factors maintain cell viability and promote cell growth by stimulating glucose transport into cells and by progressing cells through the cell cycle. In the short term, effects on glucose transport involve transporter activation, while in the longer term increased gene expression is involved. This study aimed to investigate growth factor regulation of glucose transport in an interleukin (IL)-3-dependent bone marrow-derived cell line and its oncogene-transformed counterparts. 32D clone 3 (32Dcl3) cells and cells transfected with temperature-sensitive (ts) ras and abl oncogenes, were treated with and without IL-3 and their ability to take up 2-deoxy-d-glucose compared. Transformed cells, which are not dependent on IL-3 for growth at the permissive temperature of 32 °C, exhibited a two- to six-fold higher proliferative response, enhanced tyrosine kinase activity and c-myc expression than control cells optimally stimulated with IL-3. Compared with control 32Dcl3 cells, 2-deoxy-d-glucose uptake was also 36–76% higher in transformed cells. The increased glucose uptake in transformed cells was consistent with 2.5-fold higher affinity of the glucose transporters for glucose. IL-3 stimulated glucose uptake in both control and oncogene-transformed cells. With control and ras-transformed cells, enhanced glucose uptake in response to IL-3 was associated with increased affinity of glucose transporters for glucose but with abl-transformed cells, no significant affinity changes were observed. IL-3 also increased glucose transporter expression in both control and oncogene-transformed cells, suggesting that increased transporter expression as well as changes in transporter affinity for glucose can affect glucose uptake.

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The hypoxic tumor microenvironment is associated with malignant progression and poor treatment response. The glucose transporter Glut-1 is a prognostic factor and putative hypoxia marker. So far, studies of Glut-1 in cancer have utilized conventional immunohistochemical analysis in a series of individual biopsy or surgical specimens. Tissue microarrays, however, provide a rapid, inexpensive means of profiling biomarker expression. To evaluate hypoxia markers, tissue cores must show the architectural features of hypoxia; i.e. viable tissue surrounding necrotic regions. Glut-1 may be a useful biomarker to validate tissue microarrays for use in studies of hypoxia-regulated genes in cancer. In this study, we carried out immunohistochemical detection of Glut-1 protein in many tumor and normal tissue types in a range of tissue microarrays. Glut-1 was frequently found in peri-necrotic regions, occurring in 9/34 lymphomas, 6/12 melanomas, and 5/16 glioblastomas; and in 43/54 lung, 22/84 colon, and 23/60 ovarian tumors. Expression was rare in breast (6/40) and prostate (1/57) tumors, and in normal tissue, was restricted to spleen, tongue, and CNS endothelium. In conclusion, tissue microarrays enable the observation of Glut-1 expression in peri-necrotic regions, which may be linked to hypoxia, and reflect previous studies showing differential Glut-1 expression across tumor types and non-malignant tissue.
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The aim of this study was to investigate the correlations between ¹⁸F-FDG uptake by bone marrow and various hematological parameters.
Forty-eight patients who underwent FDG-PET/CT studies and also received hematological examinations within 5 days before or after the PET study were included in this study. All patients had not received chemotherapy. FDG uptake by bone marrow was measured as a standardized uptake value (SUV) on three-dimensional PET/CT fusion images, and the uptake ratio (UR) of the SUV of bone marrow to the SUV of longitudinal dorsal muscle was calculated. The correlations between the SUV and the UR of bone marrow and various hematological parameters were evaluated.
Bone marrow FDG uptake was strongly correlated with the white blood cell counts but was not significantly correlated with the red blood cell and platelet counts. The neutrophil count was significantly correlated with bone marrow FDG uptake but the lymphocyte count was not.
FDG uptake by bone marrow was specifically correlated with the neutrophil count, suggesting that the FDG uptake by bone marrow reflects marrow metabolism that is mainly regulated by granulocyte progenitors and stimulated by endogenous hematopoietic growth factors. They may also be helpful in interpreting PET images, especially for diagnosing bone marrow involvement by malignancy.
Activation of the PI3K/mTOR pathway by BCR-ABL contributes to increased production of reactive oxygen species
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Citation Excerpt :
Phosphorylation of 2-DOG to 2-DOG-6-phosphate reduces the amount of intracellular glucose-6-phosphate formation, which in turn has recently been suggested to be involved in the regulation of ROS levels.13 In cells transformed by the BCR-ABL or v-Abl tyrosine kinases, the uptake of glucose through the glucose transporter is tightly regulated by the oncoprotein itself.14-16 Activated ABL kinases are likely to regulate glucose uptake by increasing the affinity of glucose to Glut1 and regulating transporter activity, but there may be additional mechanisms.17-19
BCR-ABL oncoprotein-expressing cells are associated with a relative increase of intracellular reactive oxygen species (ROS), which is thought to play a role in transformation. Elevated ROS levels in BCR-ABL-transformed cells were found to be blocked by the mitochondrial complex I inhibitor rotenone as well as the glucose transport inhibitor phloretin, suggesting that the source of increased ROS might be related to increased glucose metabolism. The glucose analog 2-deoxyglucose (2-DOG) reduced ROS to levels found in non-BCR-ABL-transformed cells and inhibited cell growth alone or in cooperation with imatinib mesylate (Gleevec). A mutant of BCR-ABL that is defective in transformation of myeloid cells, Tyr177Phe, was also found to be defective in raising intracellular ROS levels. Glucose metabolism in BCR-ABL-transformed cells is likely to be mediated by activation of the phosphatidylinositol-3′-kinase (PI3K) pathway, which is regulated through this site. Inhibition of PI3K or mTOR led to a significant decrease in ROS levels. Overall, our results suggest that elevated levels of ROS in BCR-ABL-transformed cells are secondary to a transformation-associated increase in glucose metabolism and an overactive mitochondrial electron transport chain and is specifically regulated by PI3K. Finally, these results hint at novel targets for drug development that may aid traditional therapy. (Blood. 2005; 105:1717-1723)
Hemopoietic cell transformation is associated with failure to downregulate glucose uptake during the G2/M phase of the cell cycle
2004, Experimental Cell Research
Citation Excerpt :
Glucose uptake can be so pronounced that 18F2-deoxyglucose, a radioactive analog of glucose, is now routinely used to locate and assess tumour metabolism. Transfection of cells with a single oncogene, for example, v-src[11], H-ras[12] and v-abl[13], markedly increases glucose uptake. Transformed cells maintain high glucose uptake by both acute and chronic mechanisms including overexpression of glucose transporters [14,15], atypical expression [16] and activation of transporter subtypes [17].
Growth factors and cytokines initiate multiple signal transduction pathways that lead to cell survival, cell cycle progression or differentiation. A common feature of these pathways is increased cellular metabolism and glucose uptake. Furthermore, the energy requirements of many cancers and transformed cell lines are met by constitutive upregulation of glucose uptake. Relationships among transforming events, glucose uptake and cell cycle progression are not well understood. Here we investigated the regulation of glucose transport during the cell cycle of growth factor-dependent 32D cells, primary T-cells, src-transformed 32D cells and Jurkat cells. Cells were enriched in the G1, S and G2/M phases of the cell cycle, and glucose transporter expression and 2-deoxyglucose uptake were measured. Glucose transporter expression increased with cell volume as cells progressed through the cell cycle. Growth factor-dependent 32D cells and T-lymphocytes were characterised by increased 2-deoxyglucose uptake from G1 to S and reduced uptake at G2/M, with the highest specific activity of transporters in the S phase. In contrast, src-transformed 32D cells and Jurkat cells showed increased 2-deoxyglucose uptake from S to G2/M, with the highest glucose transporter specific activity in G2/M. Our results show that glucose transport is regulated in a cell cycle-dependent manner and suggest that this regulation may be altered in transformed cells.
Mechanisms of basal and cytokine-induced uptake of glucose in normal human eosinophils: Relation to apoptosis
2003, Respiratory Medicine
A link between glucose transport and apoptosis was suggested. We studied the mechanisms of glucose transport in human eosinophils by means of the uptake of the positron emitting analogue, ¹⁸Fluoro-2-Deoxyglucose (FDG) and apoptosis by means of flow cytometry. FDG uptake was inhibited by antibodies to GLUT1, 3 and 4 and by cytochalasin B. The anti-apoptotic principles IL-5, GM-CSF, IL-3 enhanced the uptake, whereas the apoptosis-inducing principles anti-CD95 (anti-Fas) and exposure to serum-coated Sephadex particles caused a reduction. Also TNF-α enhanced the uptake. Other cytokines such as IL-2, IL-4, IL-8, RANTES and MCP-3 had no effect on the glucose uptake. 2-Deoxyglucose, antibodies to GLUT4 and CD95 induced apoptosis. The basal FDG-uptake was unaffected by PKC inhibitors Ro-31-8220, Gö-6983 and Gö-6976, whereas the latter inhibited the IL-5-enhanced uptake possibly due to the inhibition of PKCμ. Protein tyrosine kinase and PI-3 kinase inhibitors inhibited IL-5-enhanced FDG-uptake only. In contrast MEK inhibitors inhibited the basal uptake only. Inhibitors of p38 MAPkinase inhibited both basal and IL-5 enhanced uptake. We conclude that glucose uptake in eosinophils is governed by specific intracellular mechanisms involving mobilization of GLUTs, Ca²⁺ and the activation of the MAP kinase pathway and that the IL-5-enhanced uptake uniquely seems to involve PKCμ activity. Our results suggest a close link between apoptosis and glucose transport in human eosinophils.
N-glycosylation of glucose transporter-1 (Glut-1) is associated with increased transporter affinity for glucose in human leukemic cells
1999, Leukemia Research
To elucidate the role of N-glycosylation in the functional activity of the universal glucose transporter, Glut-1, we investigated effects of the N-glycosylation inhibitor, tunicamycin, on glucose transport by human leukemic cell lines K562, U937 and HL60. Treatment with tunicamycin produced a 40–50% inhibition of 2-deoxyglucose uptake and this was associated with a 2–2.5-fold decrease in transporter affinity for glucose (K_m) without a change in V_max. Leukemic K562, U937 and HL60 cells expressed Glut-1 transporter protein. With K562 cells Glut-1 appeared as a broad band of 50–60 kDa, whereas with U937 and HL60 cells a diffuse band was observed at approximately 55 kDa. Treatment of K562 cells with tunicamycin for 18 h, resulted in extensive loss of the 50–60 kDa glycoprotein, appearance of a 30–40 kDa band and increased staining of a 45 kDa band. With U937 cells, tunicamycin treatment resulted in the appearance of a 30–40 kDa band and increased staining of a 45 kDa band. With HL60 cells loss of the 55 kDa Glut-1 band was observed and a band of 45 kDa appeared. Tunicamycin-treatment resulted in 75–90% inhibition in [³H]mannose incorporation but only 20–25% inhibition in [³H]thymidine and [³H]leucine incorporation. In contrast, tunicamycin had little effect on the viability and MTT responses of the cells used. These results suggest that in leukemic cells N-glycosylation of Glut-1 plays an important role in maintaining its structure and functional integration.

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Original articleRegulation of glucose transport by interleukin-3 in growth factor-dependent and oncogene-transformed bone marrow-derived cell lines

Abstract

Cell Signalling

Cell

Blood

Methods in Enzymology

Analytical Biochemistry

Blood

IL-3 facilitates glucose transport in a myeloid cell line by regulating the affinity of the glucose transporter for glucose: involvement of protein phosphorylation in transporter activation

Biochemical Journal

IL-3 facilitates lymphocyte hexose transport by enhancing the intrinsic activity of the transport system

International Immunology

On the origins of cancer cells

Science

Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes

Science

Temperature-dependent alterations in sugar transport in cells infected by a temperature-sensitive mutant of Rous sarcoma virus

Transformation of rat fibroblasts by FSV rapidly increases glucose transporter gene transcription

Science

Original article
Regulation of glucose transport by interleukin-3 in growth factor-dependent and oncogene-transformed bone marrow-derived cell lines